Organism (including strain): Influenza A virus (strain A/Tokyo/3/1967 H2N2)
Etiologic Risk Group: 2
Disease Information:
Influenza virus is frequently changing; a new strain continues to appear and pose a threat to public health. Two molecules cover the surface of the virus that are a major contributor to its infectivity: hemagglutinin and neuraminidase. Hemagglutinin plays a major as the virus approaches a cell, binding to polysaccharide chains on the cell surface and then injecting the viral genome into the cell. Conversely, neuraminidase functions mainly when the virus leaves an infected cell. It ensures that the virus does not remain on the cell surface by clipping off the ends of membrane polysaccharide chains.
Structure (PDB or Homology Model): 1NN2 (LINK: https://www.rcsb.org/structure/1nn2) Similarity to Human Structures: No similar human structures were found via BLASTP.
Purification Method:
50 ml anti-FLAG affinity column (Mini-Leak Low; Kem-En-Tec A/S, Copenhagen, Denmark)
Protein would be eluted with TBA buffer containing FLAG peptide
Image of Protein:
Amino Acid Sequence:
VEYRNWSKPQCQITGFAPFSKDNSIRLSAGGDIWVTREPYVSCDPVKCYQFALGQGTTLDNKHSNDTVHDRIPHRTLLMNELGVPFHLGTRQVCIAWSSSSCHDGKAWLHVCITGDDKNATASFIYDGRLVDSIGSWSQNILRTQESECVCINGTCTVVMTDGSASGRADTRILFIEEGKIVHISPLAGSAQHVEECSCYPRYPGVRCICRDNWKGSNRPVVDINMEDYSIDSSYVCSGLVGDTPRNDDRSSNSNCRDPNNERGTQGVKGWAFDNGNDLWMGRTISKDLRSGYETFKVIGGWSTPNSKSQINRQVIVDSDNRSGYSGIFSVEGKSCINRCFYVELIRGRKQETRVWWTSNSIVVFCGTSGTYGTGSWPDGANINFMPI Length of Protein in Amino Acids: 388aa
NCBI Gene# or RefSeq#: GenBank: AAO46245
Reference Sequence: YP_308872.1
PDB ID: 1NN2
Organism (including strain): Influenza A virus (strain A/Tokyo/3/1967 H2N2)
Etiologic Risk Group: 2
Disease Information:
Influenza virus is frequently changing; a new strain continues to appear and pose a threat to public health. Two molecules cover the surface of the virus that are a major contributor to its infectivity: hemagglutinin and neuraminidase. Hemagglutinin plays a major as the virus approaches a cell, binding to polysaccharide chains on the cell surface and then injecting the viral genome into the cell. Conversely, neuraminidase functions mainly when the virus leaves an infected cell. It ensures that the virus does not remain on the cell surface by clipping off the ends of membrane polysaccharide chains.
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.):
https://www.ncbi.nlm.nih.gov/protein/AAO46245?report=genbank&log$=protalign&blast_rank=1&RID=E505869S014
Essentiality of Protein: This protein catalyzes the release of viruses from the host cell, essential for influenza virus replication.
Complex of Proteins: no
Druggable Target:
Protein is known to be druggable. Listed below are two drugs that mimic the natural substrate of the enzyme:
EC#: 3.2.1.18
Brenda EC# Page: https://www.brenda-enzymes.org/enzyme.php?ecno=3.2.1.18&Suchword=&reference=&UniProtAcc=&organism%5B%5D=influenza+A+virus+A%2Ftokyo%2F3%2F67&show_tm=0
BRENDA enzyme mechanism schematic:
Enzyme Assay Information:
Sialic Acid (NANA) Assay Protocol Information
- Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
- LINK: http://www.abcam.com/sialic-acid-nana-assay-kit-ab83375.html
Cost and Quantity of Reagents:Structure (PDB or Homology Model): 1NN2 (LINK: https://www.rcsb.org/structure/1nn2)
Similarity to Human Structures: No similar human structures were found via BLASTP.
Current Inhibitors:
- 2,3-dehydro-2-sialic acid – 70.2% at 0.1 mM
- o https://www.brenda-enzymes.org/enzyme.php?ecno=3.2.1.18
- 2-deoxy 2,3-dehydro-N-acetyl
- o https://www.ncbi.nlm.nih.gov/pubmed/8371267
Purification Method:50 ml anti-FLAG affinity column (Mini-Leak Low; Kem-En-Tec A/S, Copenhagen, Denmark)
Protein would be eluted with TBA buffer containing FLAG peptide
Image of Protein:
Amino Acid Sequence:
VEYRNWSKPQCQITGFAPFSKDNSIRLSAGGDIWVTREPYVSCDPVKCYQFALGQGTTLDNKHSNDTVHDRIPHRTLLMNELGVPFHLGTRQVCIAWSSSSCHDGKAWLHVCITGDDKNATASFIYDGRLVDSIGSWSQNILRTQESECVCINGTCTVVMTDGSASGRADTRILFIEEGKIVHISPLAGSAQHVEECSCYPRYPGVRCICRDNWKGSNRPVVDINMEDYSIDSSYVCSGLVGDTPRNDDRSSNSNCRDPNNERGTQGVKGWAFDNGNDLWMGRTISKDLRSGYETFKVIGGWSTPNSKSQINRQVIVDSDNRSGYSGIFSVEGKSCINRCFYVELIRGRKQETRVWWTSNSIVVFCGTSGTYGTGSWPDGANINFMPI
Length of Protein in Amino Acids: 388aa
Molecular Weight: 43093.06 Daltons
Molar Extinction Coefficient (with all Cys residues reduced): 83880
TMpred graph Image:
LINK: https://embnet.vital-it.ch/cgi-bin/TMPRED_form_parser
CDS Gene Sequence:
agcaaaagca ggagtgaaaa tgaatccaaa tcaaaagata ataacaattg gctctgtctc tctcaccatt gcaacagtat gctttctcat gcagattgcc atcttggtaa ctactgtaac attgcacttt aagcaacatg agtgcgactc ccccgcgagc aaccaagtaa tgccgtgtga accaataata atagaaagga acataacaga gatagtgtat ttgaataaca ccaccataga gaaagagata tgccccaaag tagtggaata cagaaattgg tcaaagccgc aatgtcaaat tacaggattt gcaccttttt ctaaggacaa ttcaatccgg ctttctgctg gtggggacat ttgggtgacg agagaacctt atgtgtcatg cgatcctgtc aagtgttatc aatttgcact cgggcagggg accacactag acaacaaaca ttcaaatgac acagtacatg atagaatccc tcatcgaacc ctattaatga atgagttggg tgttccattt cacttaggaa ccaggcaagt gtgtatagca tggtccagct caagttgtca cgatggaaaa gcatggctgc atgtttgtat cactggggat gataaaaatg caactgctag cttcatttat gacgggaggc ttgtggacag tattggttca tggtctcaaa atatcctcag aacccaggag tcggaatgcg tttgtatcaa tgggacttgc acagtagtaa tgactgatgg aagtgcttca ggaagagccg atactagaat actattcatt gaagagggga aaattgtcca tattagccca ttggcaggaa gtgctcagca tgtagaggag tgttcctgtt atcctcgata tcctggcgtc agatgtatct gcagagacaa ctggaaaggc tctaataggc ccgtcgtaga cataaatatg gaagattata gcattgattc cagttatgtg tgctcagggc ttgttggcga cacacctaga aacgatgaca gatctagcaa tagcaattgc aggaatccta acaatgagag agggactcaa ggagtgaaag gctgggcctt tgacaatgga aatgacttgt ggatgggaag aacaatcagc aaggatttac gctcaggtta tgaaactttc aaagtcattg gtggttggtc cacacctaat tccaaatcgc agatcaatag acaagtcata gttgacagtg ataatcggtc aggttactct ggtattttct ctgttgaggg caaaagctgc atcaataggt gcttttatgt ggagttgata aggggaagga aacaggagac tagagtatgg tggacctcaa acagtattgt tgtgttttgt ggcacttcag gtacctatgg aacaggctca tggcctgatg gggcgaacat caatttcatg cctatataag ctttcgcaat tttagaaaaa aactccttgt ttctact